WO2021139642A1 - 一种细胞程序性死亡受体1抗体制剂及其用途 - Google Patents

一种细胞程序性死亡受体1抗体制剂及其用途 Download PDF

Info

Publication number
WO2021139642A1
WO2021139642A1 PCT/CN2021/070248 CN2021070248W WO2021139642A1 WO 2021139642 A1 WO2021139642 A1 WO 2021139642A1 CN 2021070248 W CN2021070248 W CN 2021070248W WO 2021139642 A1 WO2021139642 A1 WO 2021139642A1
Authority
WO
WIPO (PCT)
Prior art keywords
mass
pharmaceutical preparation
monoclonal antibody
volume
content
Prior art date
Application number
PCT/CN2021/070248
Other languages
English (en)
French (fr)
Inventor
方源
韩冬梅
Original Assignee
上海复宏汉霖生物技术股份有限公司
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 上海复宏汉霖生物技术股份有限公司 filed Critical 上海复宏汉霖生物技术股份有限公司
Priority to JP2022541225A priority Critical patent/JP2023510229A/ja
Priority to AU2021205538A priority patent/AU2021205538A1/en
Priority to KR1020227026697A priority patent/KR20220124208A/ko
Priority to EP21738991.5A priority patent/EP4088740A4/en
Priority to CA3162976A priority patent/CA3162976A1/en
Priority to US17/758,399 priority patent/US20230048612A1/en
Priority to BR112022012768A priority patent/BR112022012768A2/pt
Publication of WO2021139642A1 publication Critical patent/WO2021139642A1/zh
Priority to ZA2022/07051A priority patent/ZA202207051B/en

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
    • C07K16/2818—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against CD28 or CD152
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A61K39/39591—Stabilisation, fragmentation
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/02—Inorganic compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
    • A61K47/12—Carboxylic acids; Salts or anhydrides thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
    • A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
    • A61K47/183—Amino acids, e.g. glycine, EDTA or aspartame
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/08—Solutions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/94—Stability, e.g. half-life, pH, temperature or enzyme-resistance

Definitions

  • the invention relates to the field of biotechnology, in particular to a programmed cell death receptor 1 (PD-1) antibody preparation and its use.
  • PD-1 programmed cell death receptor 1
  • PD-1 Programmed death receptor 1
  • CD279 is an important immunosuppressive molecule, a member of the CD28 family of immunoglobulin superfamily, and a type I transmembrane with a molecular weight of 50-55kD Glycoprotein.
  • PD-1 is continuously expressed on the surface of T cells, B cells, natural killer cells, monocytes and bone marrow cells.
  • PD-1 has two known ligands, PD-L1 and PD-L2.
  • studies have found that the combination of PD-1 and its ligand PD-L1 can transmit inhibitory signals and reduce the proliferation of CD8+ T cells in lymph nodes, and PD-1 can also control the antigens in lymph nodes by regulating the Bcl-2 gene.
  • Protein (monoclonal antibody, etc.) pharmaceutical preparations are suitable for parenteral administration, including intravenous, intramuscular, intraperitoneal or subcutaneous injection.
  • the liquid preparation is convenient to use, but the stability is poor, and the storage conditions are high. Generally, it needs to be stored under low temperature conditions.
  • the protein in the liquid preparation is easy to form aggregates or particles, which will also cause the stability of the monoclonal antibody preparation to decrease.
  • the proteins contained, especially monoclonal antibody drugs are difficult to maintain good physical stability, chemical stability and biological stability during the storage period, especially under non-refrigerated conditions. Therefore, it is necessary to study against such drugs. Stable buffer system for drugs.
  • stabilizers that can be added to protein (mab) drugs include surfactants, sugars and polyols, salts, amino acids/amino acid salts, and some macromolecular compounds.
  • antibody drugs have different structural properties and are easy to polymerize. At present, there is still a lack of stable protein preparations that meet the needs of the pharmaceutical industry.
  • One of the objectives of the present invention is to provide a pharmaceutical preparation containing an anti-PD-1 monoclonal antibody.
  • the present invention firstly determines the use of citrate-sodium citrate buffer as the preferred buffer system for anti-PD-1 monoclonal antibody drug preparations through screening and formulation optimization research.
  • the pharmaceutical preparation includes an anti-PD-1 monoclonal antibody, a buffer, a stabilizer, and a surfactant, and optionally an isotonicity regulator.
  • the pharmaceutical preparation includes: anti-PD-1 monoclonal antibody, citrate-sodium citrate buffer, protein protectant and surfactant.
  • the content of the anti-PD-1 monoclonal antibody in the pharmaceutical preparation is preferably 5-50 mg/mL, more preferably 8-30 mg/mL, preferably 10.0 mg/mL.
  • the anti-PD-1 monoclonal antibody optionally has a heavy chain variable region shown in SEQ ID NO. 7 and a light chain variable region shown in SEQ ID NO. 8.
  • the anti-PD-1 monoclonal antibody is an h1G4 antibody, which comprises the heavy chain shown in SEQ ID NO.10 and the light chain shown in SEQ ID NO.9.
  • the pharmaceutical preparation contains 10 mg/ml of an anti-PD-1 monoclonal antibody, wherein the anti-PD-1 monoclonal antibody is an h1G4 antibody, comprising the heavy chain and SEQ ID NO.10 shown in SEQ ID NO.10. The light chain shown in ID NO.9.
  • the concentration of the citrate-sodium citrate is preferably 10-50 mM, more preferably 15-30 mM, and preferably 20 mM. In one embodiment, the content of the citrate-sodium citrate buffer is 10-30 mM.
  • the pharmaceutical preparation contains 20 mmol/L citrate-sodium citrate buffer.
  • the pH value of the pharmaceutical preparation is preferably pH 5.0-6.5.
  • the pH value can be 5.0, 5.5, 6.0, or 6.5.
  • the pH value of the pharmaceutical preparation is pH 5.5.
  • the preparation of the present invention also adds a protein protective agent to provide product stability, selected from conventional protein protective agents in the art, which can protect the stability of protein drugs and protect the function of protein drugs from changes in conditions (for example, : Freezing, freeze-drying or other changes in preparation conditions).
  • Protein protective agents preferably include: carbohydrates, proteins, amino acids, polymers, salts, amines, surfactants and the like. More preferably, the protein protective agent includes one or more of sucrose, mannitol or glycine. In one embodiment, the protein protective agent is selected from one or more of sucrose, glycine and mannitol. In a specific embodiment, the protein protective agent is mannitol, sucrose or glycine.
  • the content of the protein protective agent is preferably 0.5-5% by mass/volume percentage (g/L). In another embodiment, the protein protectant content is 1 to 5% by mass/volume, wherein the mass/volume ratio is g/L.
  • the protein protective agent is selected from one or more of sucrose, glycine and mannitol, and its content is 0.5-5% by mass/volume, wherein the mass/volume ratio is g/L.
  • the pharmaceutical preparation contains sucrose in a mass/volume percentage of 1-3%. In another embodiment, the pharmaceutical preparation contains 1% glycine in a mass/volume percentage. In a preferred embodiment, the pharmaceutical preparation contains 3% mannitol in a mass/volume percentage. The mass/volume ratio mentioned above is g/L.
  • the preparation of the present invention may include a surfactant, selected from conventional surfactants in the art, preferably nonionic surfactants.
  • surfactants herein preferably include: polysorbates (e.g., polysorbate 20 and polysorbate 80); poloxamers (e.g., poloxamer 188); Triton; sodium lauryl sulfate (SDS); sodium lauryl sulfate; sodium octyl glycoside; lauryl-, myristyl-, linoleyl-, or stearoyl-sulfobetaine; lauryl-, myristyl-, linoleyl -, or stearoyl-sarcosine; linoleyl-, myristyl, or cetyl-betaine; lauroamidopropyl-, cocoamidopropyl-, linoleamidopropyl- , Myristoylaminopropyl-, palmitoylaminopropyl-, or isostearamidopropyl-betaine (such as lauroamidopropyl
  • the content of the surfactant is preferably 0.01%-0.05% by volume. In a preferred embodiment, the content of the surfactant is 0.02% by volume.
  • the pharmaceutical preparation contains 0.02% polysorbate 80 by volume.
  • the pharmaceutical preparation includes: anti-PD-1 monoclonal antibody, citrate-sodium citrate buffer, protein protectant and surfactant, wherein
  • the content of the citrate-sodium citrate buffer is 10-30 mM;
  • the protein protective agent is selected from one or more of sucrose, glycine and mannitol, and the content is 0.5-5% by mass/volume, wherein the mass/volume percentage is g/L;
  • the surfactant is polysorbate 80, and the content is 0.01%-0.05% by volume;
  • the anti-PD-1 monoclonal antibody has a heavy chain variable region shown in SEQ ID NO. 7 and a light chain variable region shown in SEQ ID NO. 8.
  • the formulation of the present invention may or may not contain an isotonic adjusting agent as required, wherein the isotonic adjusting agent is a conventional isotonic adjusting agent in the art.
  • the isotonicity adjusting agent is sodium chloride.
  • the content of the isotonicity adjusting agent is 0.1-0.5% by mass/volume, wherein the mass/volume ratio is g/L.
  • the pharmaceutical preparation contains sodium chloride with a mass/volume percentage of 0.3%, wherein the mass/volume ratio is g/L.
  • the present invention carries out a high temperature accelerated test at 40°C for a maximum of 4 weeks for each component of the formulation to detect and evaluate the stability data of each formulation.
  • the test items include: appearance, concentration, turbidity (A350), pH value, penetration Pressure, purity (SEC-HPLC, CEX-HPLC) and other tests. Combining the test results, analyze the optimal formulation formula.
  • the pharmaceutical preparation contains: anti-PD-1 monoclonal antibody 10 mg/ml, 20 mmol/L citrate-sodium citrate buffer, 3% mannitol by mass/volume percentage, volume The percentage is 0.02% polysorbate 80, the mass/volume percentage is 0.3% sodium chloride, the pH is 5.5, and the mass/volume ratio is g/L.
  • the pharmaceutical preparation contains: anti-PD-1 monoclonal antibody 10 mg/ml, 20 mmol/L citrate-sodium citrate buffer, 1% mass/volume glycine, volume The percentage is 0.02% polysorbate 80, the mass/volume percentage is 0.3% sodium chloride, the pH is 5.5, and the mass/volume ratio is g/L.
  • the dosage form of the pharmaceutical preparation may be a conventional dosage form in the art, and preferably includes a liquid preparation for injection or a freeze-dried preparation, and the like.
  • the liquid preparations for injection preferably include subcutaneous injection preparations, intravenous injection preparations, intraperitoneal administration preparations, intramuscular injection preparations, intravenous/subcutaneous injection preparations or vitreous injection preparations, and the like.
  • the liquid preparation for injection preferably includes a water needle injection preparation, a prefilled needle injection preparation, etc., preferably a water needle injection preparation, which can be used for intravenous injection. Therefore, in another aspect, the present invention also relates to the application of the pharmaceutical preparation in the preparation of a liquid preparation for injection or a lyophilized preparation.
  • any numerical value such as the concentration, concentration range, or content described herein, should be understood to be modified by the term "about” in any case. Therefore, the value usually includes ⁇ 10% of the stated value. For example, a concentration of 1 mg/mL includes 0.9 mg/mL to 1.1 mg/mL. Likewise, the concentration range of 1% to 10% (w/v) includes 0.9% (w/v) to 11% (w/v). As used herein, unless the context clearly dictates otherwise, the use of a range of values explicitly includes all possible subranges, all individual values within the range, including integers and fractions of values within the range.
  • composition optionally comprising an isotonicity adjusting agent encompasses the inclusion or absence of an isotonicity adjusting agent.
  • pharmaceutical preparation refers to a medicine administered to a patient in need of treatment, which can be in solid or liquid forms (ie, "dosage form"), such as liquid preparations (for example, liquid preparations for injection), powders, and freeze-dried preparations.
  • the pharmaceutical preparation of the present invention has undergone high-temperature accelerated testing, and the main detection indicators have no obvious changes; after 4 weeks of accelerated processing under the condition of 40°C, the test shows that the main detection indicators have no significant changes, and are relatively stable. The results all meet the requirements of the current quality standards of this product, showing good stability.
  • FIG. 1 Buffer system screening study of h1G4 antibody preparations. Accelerated stability test (40°C) turbidity histogram.
  • Reagent sources the reagents involved in the experimental part of the present invention are all commercially available products.
  • the pipette draws 20 ⁇ l of sample and 2 osmolarity standards of 290mOsmol/kg, and uses Advanced 2020 osmometer to determine the osmolarity of the samples and standards.
  • the samples were analyzed using Agilent 1260 High Performance Liquid Chromatography, TOSOH TSK G3000 (300*7.8mm) chromatographic column, the column temperature was room temperature, the flow rate was 0.5mL/min, and the detection wavelength was 280nm.
  • the mobile phase composition is 100mM PB, 0.5% NaCl, pH 6.8, the test sample is diluted to about 1.0mg/mL with the mobile phase, 50 ⁇ L is injected, and the stop time is 30min.
  • the samples were analyzed by Agilent 1260 High Performance Liquid Chromatography, Thermo ProPac WCX-10 column (4 ⁇ 250mm), column temperature was 35°C, flow rate was 1.0 mL/min, detection wavelength was 280 nm, and the mobile phase elution gradient is shown in Table 1.
  • the composition of mobile phase A is CX-1 Gradient buffer A, pH 5.6
  • the composition of mobile phase B is CX-1 Gradient buffer B, pH 10.2.
  • the test sample is diluted to about 1.0 mg/mL with mobile phase A, and 20 ⁇ L is injected.
  • the sample needs to be diluted to 1.0 mg/ml with placebo before scanning.
  • the PD-1 antibody h1G4 is derived from the h1G4 disclosed in the patent application WO2018052818, and its preparation method is fully cited in WO2018052818.
  • the amino acid sequence of PD-1 antibody h1G4 is as follows:
  • h1G4 antibody heavy chain (HC) SEQ ID NO: 10
  • B1 ⁇ B6 two different formulation buffer systems were selected as alternative formulations (hereinafter referred to as B1 ⁇ B6), and the target drug h1G4 antibody concentration was 10.0 mg/mL.
  • B1 ⁇ B3 are 20mmol/L citrate-sodium citrate buffer system, with pH values of 5.0, 5.5 and 6.0 respectively
  • B4 ⁇ B6 are 20mmol/L histidine-histidine hydrochloric acid buffer system, pH value Respectively 5.0, 5.5 and 6.0
  • the above-mentioned buffer system contains 3% mannitol, 0.02% polysorbate 80 and 0.3% sodium chloride as auxiliary materials, and the percentage of the concentration of auxiliary materials refers to the mass/volume ratio (w/v, g/L). See Table 3 for alternative prescription information for the screening study of h1G4 antibody preparation buffer system.
  • a 0.22 ⁇ m disposable sterile filter is used in the biological safety cabinet to filter the samples, aseptically dispensed into 2mL vials, add a 13mm rubber stopper, and roll a 13mm aluminum-plastic composite cover. Place the aliquoted sample in a constant temperature medicinal storage box at 40°C and 2-8°C for 2 weeks. Take samples at the 0th, 1st, and 2nd weeks for testing and analysis. The test items are shown in Table 4.
  • the other buffer systems are colorless and micro opalescent liquid with no obvious visible foreign matter.
  • the concentration is in the range of 9-11mg/mL.
  • the molarity is 300 ⁇ 340mOsmol/kg.
  • the main peak content of SEC is greater than 99.0%, and the aggregate content is 0.6-0.7%.
  • the CEX main peak content is 47.0-48.6%, the acidic peak content is 11.0-14.3%, the basic peak content is 38.6-40.9%, and the R-CE-SDS purity is 94.1-96.9%.
  • the basic physical and chemical test results showed that at week 0, the quality of each buffer system was good, and there was no significant difference (see Table 5).
  • the protein concentration of all citrate-sodium citrate buffer systems did not change significantly, all within the range of 9-11mg/mL, but the histidine-histidine hydrochloric acid buffer system
  • the concentration in C50, C60 and H55 buffer system is significantly increased, and protein precipitation occurs in the C50, C60 and H55 buffer systems.
  • the turbidity of the histidine-histidine hydrochloride buffer system (A350: Use NanoDrop 2000C, add 2.5 ⁇ L, and measure the sample at 350nm wavelength The absorbance value below.
  • the SEC main peak content of the citrate-sodium citrate buffer system decreased by 0.1-2.5%, the aggregate content increased by 0.1-0.4%, and the fragment content increased by 0-2.1%.
  • all histidine-histidine hydrochloride buffer systems have obvious degradation reactions, so the citrate-sodium citrate buffer system is better than histidine-histidine hydrochloride system, and the optimal pH is 5.5.
  • N/A means not applicable.
  • the target protein h1G4 antibody concentration is 10.0 mg/mL.
  • the buffer system is a 20mmol/L citric acid-sodium citrate solution with a pH of 5.5. Except that prescription F3 does not contain sodium chloride, the other prescriptions all contain 0.3% sodium chloride and 0.02% polysorbate 80.
  • the content of mannitol in prescriptions F1, F2, and F4 is 1%, 3%, and 5%, respectively.
  • the formulations of F2, F5, and F6 contain isotonic 3% mannitol, 3% sucrose and 1% glycine, respectively, to screen the optimal excipient system that contributes to the long-term stability of h1G4 antibody preparations. See Table 6 for alternative prescription information for the screening study of h1G4 antibody preparation excipients.
  • h1G4 antibody preparation excipient screening study single factor test design 6 prescriptions according to the results of the accelerated stability test at 40°C and the ranking of the excipients (see Table 7), the prescription without sodium chloride (F3) and the one containing 5% mannitol
  • the formulation (F4) is slightly less stable, and there is no significant difference between the formulation containing 1% mannitol (F1) and 3% mannitol (F2) and the formulation containing 3% sucrose (F5).
  • N/A means not applicable. *Indicating that the opalescence of F4 prescription is slightly heavier than other prescriptions after being placed at 40 ⁇ 2°C for 2 weeks.
  • formulations were screened through a single factor test, and 9 alternative formulations were designed (hereinafter referred to as F7 to F15), and the target drug h1G4 antibody concentration was 10.0 mg/mL.
  • the buffer systems for formulations F7 to F14 are all 20mmol/L pH 5.5 citrate-sodium citrate, and F15 is 20mmol/L sodium citrate-disodium hydrogen phosphate.
  • the alternative prescription information for the screening study of preparation excipients is shown in Table 8. .
  • the concentration was in the range of 9-11 mg/mL, the osmotic pressure was 185-540 mOsmol/kg, and the turbidity (A350) was 0.006-0.009.
  • the main peak content of SEC is 98.0-98.2%, and the aggregate content is 1.7-1.9%.
  • the CEX main peak content is 35.4-39.1%, the acidic peak content is 31.6-33.8%, and the basic peak content is 27.5-30.8% (see Table 10).
  • the basic physical and chemical test results showed that at week 0, the quality of each prescription was good, with no obvious difference.
  • the sodium citrate-disodium hydrogen phosphate buffer system is not suitable for the stable storage of h1G4 antibodies.
  • N/A means not applicable.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Epidemiology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Veterinary Medicine (AREA)
  • Engineering & Computer Science (AREA)
  • Immunology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Organic Chemistry (AREA)
  • Molecular Biology (AREA)
  • Biochemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • Biophysics (AREA)
  • Inorganic Chemistry (AREA)
  • Mycology (AREA)
  • Microbiology (AREA)
  • Dermatology (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Medicinal Preparation (AREA)
  • Peptides Or Proteins (AREA)

Abstract

本发明提供了一种包含抗PD-1单克隆抗体的药物制剂，其包括：抗PD-1单克隆抗体、枸橼酸-枸橼酸钠缓冲液、蛋白保护剂、表面活性剂和等渗调节剂。本发明还涉及所述的药物制剂在制备注射用液体制剂或冻干制剂中的应用。

Description

一种细胞程序性死亡受体1抗体制剂及其用途
本申请要求2020年1月6日提交的，题为“一种细胞程序性死亡受体1抗体制剂及其用途”的第202010008058.6号中国专利申请的优先权，该申请的内容整体援引加入本文。
技术领域
本发明涉及生物技术领域，具体涉及一种细胞程序性死亡受体1(PD-1)抗体制剂及其用途。
背景技术
细胞程序性死亡受体1(programmed death 1，PD-1)，也称CD279，是一种重要的免疫抑制分子，属于免疫球蛋白超家族CD28家族成员，分子量为50～55kD的I型跨膜糖蛋白。PD-1持续性表达在T细胞、B细胞、自然杀伤细胞、单核细胞以及骨髓细胞的表面。PD-1具有两个已知配体，PD-L1和PD-L2。目前，研究发现，PD-1与其配体PD-L1结合，可以传导抑制性的信号，减低淋巴结CD8+T细胞的增生，而且PD-1还可以借由调节Bcl-2基因，控制淋巴结中抗原特异性T细胞的聚积。而阻断PD-1与PD-L1的结合，可以有效阻止T淋巴细胞抑制信号的产生，从而打破免疫系统对自身组织的外周免疫耐受，促进T淋巴细胞的活化与增殖，以及细胞因子表达。因此研发新型的抗PD-1单克隆抗体，用于癌症的免疫治疗，使其具有更低的毒副作用，更佳的临床药效，成为目前的研究热点，也将给患者提供更多的药物选择。
蛋白质(单克隆抗体等)类药物制剂适合于非肠道给药，包括静脉内、肌肉内、腹膜内或皮下注射等给药方式。其液体制剂使用方便，但稳定性较差，对储存条件要求较高，一般需在低温条件下保存。此外液体制剂中蛋白易形成聚体或颗粒，也会导致单抗制剂稳定性下降。通常的药物制剂中，包含的蛋白质特别是单克隆抗体类药物，很难在储藏期内，尤其非冷藏条件下保持良好的物理稳定性、化学稳定性和生物稳定性，因此需要研究针对此类药物的稳定缓冲液体系。已有报道可在蛋白(单抗)类药物中加入的稳 定剂包括表面活性剂、糖和多元醇、盐类、氨基酸/氨基酸盐以及一些大分子化合物。但是抗体类药物的结构性质各异且易于聚合，目前还缺乏满足制药工业需求的稳定的蛋白制剂。
发明内容
本发明的目的之一是提供一种包含抗PD-1单克隆抗体的药物制剂。
本发明通过筛选和配方优化研究，首先确定使用枸橼酸-枸橼酸钠缓冲液作为抗PD-1单抗药物制剂的优选缓冲体系。具体地，该药物制剂包括抗PD-1单克隆抗体、缓冲液、稳定剂和表面活性剂、以及可选地包含等渗调节剂。
在经筛选的缓冲液体系基础上，设计单因素试验，考察蛋白质浓度、pH值以及稳定剂种类及含量、表面活性剂种类及含量、是否添加等渗调节剂等因素对蛋白稳定性的影响。从而优化筛选得出优选的其他制剂组分。
在一些实施方案中，所述药物制剂包括：抗PD-1单克隆抗体、枸橼酸-枸橼酸钠缓冲液、蛋白保护剂和表面活性剂。
所述药物制剂中抗PD-1单克隆抗体的含量较佳地为5-50mg/mL，更佳地为8-30mg/mL，优选地为10.0mg/mL。所述抗PD-1单克隆抗体可选地为具有SEQ ID NO.7所示的重链可变区和SEQ ID NO.8所示的轻链可变区。在优选的实施方案中，所述抗PD-1单克隆抗体为h1G4抗体，其包含SEQ ID NO.10所示的重链和SEQ ID NO.9所示的轻链。
在一具体实施方案中，所述药物制剂含有抗PD-1单克隆抗体10mg/ml，其中所述抗PD-1单克隆抗体是h1G4抗体，包含SEQ ID NO.10所示的重链和SEQ ID NO.9所示的轻链。
该制剂的缓冲液体系中，所述枸橼酸-枸橼酸钠的浓度较佳地为10-50mM，更佳地为15-30mM，优选地为20mM。在一实施方案中，所述枸橼酸-枸橼酸钠缓冲液含量为10-30mM。
在一具体实施方案中，所述药物制剂含有20mmol/L枸橼酸-枸橼酸钠缓冲液。
所述药物制剂的pH值较佳地为pH 5.0-6.5。例如，pH值可以为5.0、5.5、6.0或6.5。优选地，所述药物制剂的pH值为pH 5.5。
本发明制剂还添加蛋白保护剂提供产品的稳定性，选自本领域常规蛋白保护剂，所述蛋白保护剂能够保护蛋白质药物的稳定性，并保护蛋白质 药物的功能不受条件变化的影响(例如：冷冻、冻干或其他制备条件的变化)。蛋白保护剂较佳地包括：糖类、蛋白质、氨基酸、聚合物、盐、胺、表面活性剂等。更佳地，蛋白保护剂包括：蔗糖、甘露醇或甘氨酸中的一种或多种。在一实施方案中，所述蛋白保护剂选自蔗糖、甘氨酸和甘露醇中的一种或多种。在一具体实施方案中，蛋白保护剂为甘露醇、蔗糖或甘氨酸。
所述蛋白保护剂的含量较佳地为质量/体积百分比(g/L)0.5-5％。在又一实施方案中，蛋白保护剂含量为质量/体积百分比1-5％，其中质量/体积比为g/L。
在优选的实施方案中，蛋白保护剂选自蔗糖、甘氨酸和甘露醇中的一种或多种，其含量为质量/体积百分比0.5-5％，其中质量/体积比为g/L。
在一实施方案中，所述药物制剂含有质量/体积百分比为1-3％的蔗糖。在又一实施方案中，所述药物制剂含有质量/体积百分比为1％的甘氨酸。在一优选实施方案中，所述药物制剂含有质量/体积百分比为3％的甘露醇。如上所述的质量/体积比为g/L。本发明制剂可以包括表面活性剂，选自本领域常规表面活性作用剂，较佳地为非离子型表面活性剂。本文中的表面活性剂的例子较佳地包括：聚山梨酯(例如聚山梨酯20和聚山梨酯80)；泊洛沙姆(例如泊洛沙姆188)；Triton；十二烷基硫酸钠(SDS)；月桂基硫酸钠；辛基糖苷钠；月桂基-，肉豆蔻基-，亚油基-，或硬脂酰基-磺基甜菜碱；月桂基-，肉豆蔻基-，亚油基-，或硬脂酰基-肌氨酸；亚油基-，肉豆蔻基，或鲸蜡基-甜菜碱；月桂酰氨基丙基-，椰油酰氨基丙基-，亚油酰氨基丙基-，肉豆蔻酰氨基丙基-，棕榈酰氨基丙基-，或异硬脂酰氨基丙基-甜菜碱(例如月桂酰氨基丙基-甜菜碱)；肉豆蔻酰氨基丙基-，棕榈酰氨基丙基-，或异硬脂酰氨基丙基-二甲胺；甲基椰油基牛磺酸钠或甲基油基牛磺酸二钠；和MONAQUATTM系列(Mona Industries，Inc.，Paterson，N.J.)；聚乙二醇，聚丙二醇，及乙二醇和丙二醇共聚物(例如Pluronics，PF68等。在一些优选实施方案中，表面活性剂为聚山梨酯。更优选地，表面活性剂为聚山梨酯80。
所述表面活性剂的含量较佳地为体积比0.01％-0.05％。在一优选实施方案中，表面活性剂的含量为体积比0.02％。
在一具体实施方案中，所述药物制剂含有体积百分比为0.02％的聚山梨酯80。
在一具体实施方案中，所述药物制剂包括：抗PD-1单克隆抗体、枸橼 酸-枸橼酸钠缓冲液、蛋白保护剂和表面活性剂，其中
所述枸橼酸-枸橼酸钠缓冲液含量为10-30mM；
所述蛋白保护剂选自蔗糖、甘氨酸和甘露醇中的一种或多种，含量为质量/体积百分比0.5-5％，其中质量/体积百分比为g/L；
所述表面活性剂为聚山梨酯80，含量为体积比0.01％-0.05％；
所述抗PD-1单克隆抗体具有SEQ ID NO.7所示的重链可变区和SEQ ID NO.8所示的轻链可变区。
本发明的制剂中可以根据需要含有或不含有等渗调节剂，其中所述等渗调节剂为本领域常规等渗调节剂。在一优选实施方案中，等渗调节剂为氯化钠。在一实施方案中，等渗调节剂的含量为质量/体积百分比0.1-0.5％，其中质量/体积比为g/L。
在一具体实施方案中，所述药物制剂含有质量/体积百分比为0.3％的氯化钠，其中质量/体积比为g/L。
本发明对各个组分的制剂进行了40℃，最长4周的高温加速试验，检测评价各制剂配方的稳定性数据，检测项目包括：外观、浓度、浊度(A350)、pH值、渗透压、纯度(SEC-HPLC、CEX-HPLC)等测试。结合测试结果，分析得出最优的制剂配方。
在一具体实施方案中，所述药物制剂含有：抗PD-1单克隆抗体10mg/ml，20mmol/L枸橼酸-枸橼酸钠缓冲液，质量/体积百分比为3％的甘露醇，体积百分比为0.02％的聚山梨酯80，质量/体积百分比为0.3％的氯化钠，pH 5.5，其中质量/体积比为g/L。
在另一具体实施方案中，所述药物制剂含有：抗PD-1单克隆抗体10mg/ml，20mmol/L枸橼酸-枸橼酸钠缓冲液，质量/体积百分比为1-3％的蔗糖，体积百分比为0.02％的聚山梨酯80，质量/体积百分比为0.3％的氯化钠，pH 5.5，其中质量/体积比为g/L。
在又一具体实施方案中，所述药物制剂含有：抗PD-1单克隆抗体10mg/ml，20mmol/L枸橼酸-枸橼酸钠缓冲液，质量/体积百分比为1％的甘氨酸，体积百分比为0.02％的聚山梨酯80，质量/体积百分比为0.3％的氯化钠，pH 5.5，其中质量/体积比为g/L。
所述药物制剂的剂型可以为本领域常规剂型，较佳地包括注射用液体制剂或冻干制剂等等。所述注射用液体制剂较佳地包括皮下注射制剂、静脉注射制剂、腹腔给药制剂、肌肉注射制剂、静脉/皮下注射制剂或玻璃体 注射制剂等。所述注射用液体制剂较佳地包括水针注射制剂、预充针注射制剂等，优选地为水针注射制剂，该水针注射制剂可以用于静脉注射。因此，在又一方面，本发明还涉及所述药物制剂在制备注射用液体制剂或冻干制剂中的应用。
除非另有说明，本文所用的术语和短语具有下文所列的含义。特定的术语或短语在没有特别定义的情况下不应该认为是不确定的或不清楚的，而应该按照本领域技术人员通常理解的含义进行解释。当本文中出现商品名时，意在指代其对应的商品或其活性成分。本文引用的每个文件(包括所有专利、专利申请、科学出版物、制造商的说明书、指引等)，无论是上文或下文，均整体援引加入本文。本文中的任何内容均不得解释为承认本发明无法享有在先申请的优先权。
本文提供的任何和所有实施例或示例性语言(例如，“例如”)的使用仅为了更好地说明本发明，并且不对本发明的范围构成限制。
除非另有说明，否则任何数值，例如本文所述的浓度、浓度范围或含量，在任何情况下均应理解为由术语“约”修饰。因此，数值通常包括所述数值的±10％。例如，1mg/mL的浓度包括0.9mg/mL至1.1mg/mL。同样，1％至10％(w/v)的浓度范围包括0.9％(w/v)至11％(w/v)。如本文所用，除非上下文另外明确指出，否则数值范围的使用明确地包括所有可能的子范围，该范围内的所有单个数值，包括该范围内的整数以及数值的分数。
表述“包含”或与其同义的类似表述“包括”、“含有”和“具有”等是开放性的，不排除额外的未列举的元素、步骤或成分。表述“由…组成”或“由…构成”排除未指明的任何元素、步骤或成分。表述“基本上由…组成”指范围限制在指定的元素、步骤或成分，加上任选存在的不会实质上影响所要求保护的主题的基本和新的特征的元素、步骤或成分。应当理解，表述“包含”涵盖表述“基本上由…组成”和“由…组成”。
术语“可选”或“可选地”是指随后描述的事件或情况可能发生或可能不发生，该描述包括发生所述事件或情况和不发生所述事件或情况。例如，组合物可选地包含等渗调节剂涵盖包含或者不包含等渗调节剂的情况。
术语“药物制剂”指被施予需要治疗的患者的药品，其可以为固体或液体等形式(即“剂型”)，例如液体制剂(例如注射用液体制剂)、粉剂和冻干制剂等。
本发明的药物制剂经过高温加速测试，主要检测指标均无明显变化； 在40℃条件下经过4周的加速处理，经检测表明，各项主要检测指标均无明显变化，较为稳定，且所有检测结果均符合本品现行质量标准要求，表现出良好的稳定性。
附图说明
图1.h1G4抗体制剂缓冲体系筛选研究加速稳定性试验(40℃)SEC片段含量(％)柱状图。
图2.h1G4抗体制剂缓冲体系筛选研究加速稳定性试验(40℃)浊度柱状图。
具体实施方式
以下通过实施例对本发明进行进一步的详细说明，不应理解为对本发明的限制。
试剂来源：本发明的实验部分涉及的试剂均为市售产品。
实施例1.样品测试方法
1.1.渗透压检测
移液器吸取20μl样品及2份290mOsmol/kg的渗透压标准品，采用Advanced 2020型渗透压仪测定样品及标准品的渗透压值。
1.2.SEC-HPLC
采用Agilent 1260高效液相色谱，TOSOH TSK G3000(300*7.8mm)色谱柱分析样品，柱温为室温，流速0.5mL/min，检测波长280nm。流动相组成为100mM PB，0.5％NaCl，pH 6.8，供试样品用流动相稀释到1.0mg/mL左右，进样50μL，停止时间30min。
1.3.CEX-HPLC
采用Agilent 1260高效液相色谱，Thermo ProPac WCX-10色谱柱(4×250mm)分析样品，柱温为35℃，流速1.0mL/min，检测波长280nm，流动相洗脱梯度见表1所示。流动相A组成为CX-1 Gradient buffer A，pH 5.6，流动相B组成为CX-1 Gradient buffer B，pH 10.2，供试样品用流动相A稀释到1.0mg/mL左右，进样20μL。
表1.流动相洗脱梯度
时间(min) 流动相B％ 流速(mL/min)
0.00 10.0 1.0
5.00 10.0 1.0
35.00 40.0 1.0
35.01 100.0 1.0
40.00 100.0 1.0
40.01 10.0 1.0
45.00 10.0 1.0
1.4.DSC
通过差示热量扫描方法测试供试品Tm onset、Tm1及Tm2值。样品扫描前需用安慰剂稀释至1.0mg/ml。
实施例2.抗体的制备
PD-1抗体h1G4来源自专利申请WO2018052818中所公开的h1G4，其制备方法完全引用WO2018052818。PD-1抗体h1G4的氨基酸序列如下：
表2.h1G4的CDR序列
Figure PCTCN2021070248-appb-000001
Figure PCTCN2021070248-appb-000002
h1G4抗体轻链(LC)SEQ ID NO：9
Figure PCTCN2021070248-appb-000003
h1G4抗体重链(HC)SEQ ID NO：10
Figure PCTCN2021070248-appb-000004
实施例3.缓冲体系筛选研究
本研究选择了两组不同的制剂缓冲体系作为备选处方(以下分别称为B1～B6)，目标药物h1G4抗体浓度为10.0mg/mL。其中B1～B3为20mmol/L枸橼酸-枸橼酸钠缓冲体系，pH值分别为5.0、5.5及6.0；B4～B6为20mmol/L组氨酸-组氨酸盐酸缓冲体系，pH值分别为5.0、5.5及6.0；上述缓冲体系中均含有3％甘露醇、0.02％聚山梨酯80以及0.3％氯化钠作为辅料，其中辅料浓度百分比均指代质量/体积比(w/v，g/L)。h1G4抗体制剂缓冲体系筛选研究备选处方信息见表3。
表3.h1G4抗体制剂备选缓冲体系组成
Figure PCTCN2021070248-appb-000005
取适量抗体原液超滤换液，加辅料，调节浓度制备成如表3所示的处方。生物安全柜内采用0.22μm一次性无菌过滤器过滤样品，无菌分装至2mL西林瓶中，加13mm胶塞，轧13mm铝塑组合盖。将分装后样品放置于40℃及2～8℃的恒温药用保存箱内正置2周，于第0、1、2周分别取样，检测分析，测试项目详见表4。
表4.h1G4抗体缓冲体系筛选研究考察条件
Figure PCTCN2021070248-appb-000006
研究结果
高温加速试验结果
0周时除20mmol/L pH 5.0的枸橼酸体系(C50)出现颗粒外，其余缓冲体系外观均为无色微乳光液体，无明显可见异物，浓度在9～11mg/mL范围内，渗透压摩尔浓度为300～340mOsmol/kg。SEC主峰含量均大于99.0％，聚体含量为0.6～0.7％。CEX主峰含量为47.0～48.6％，酸性峰含量为11.0～14.3％，碱性峰含量为38.6～40.9％，R-CE-SDS纯度为94.1～96.9％。基础理化检测结果显示0周时，各缓冲体系质量良好，无明显差异(见表5)。
在40℃加速条件下放置2周后，所有枸橼酸-枸橼酸钠缓冲体系蛋白浓度无明显变化，均在9～11mg/mL范围内，但是组氨酸-组氨酸盐酸缓冲体系中浓度显著升高，并且C50、C60以及H55缓冲体系出现蛋白沉淀，组氨酸-组氨酸盐酸盐缓冲体系的浊度(A350：使用NanoDrop 2000C，加样2.5μL，测定样品在350nm波长下的吸光度值。重复测定三次取其均值为最终结果。)数值明显增加(见图1和图2)，故缓冲体系优劣排序为C55>C60>C50,H60>H55>H50。40±2℃条件下放置2周通过CE-SDS测定发现重链含量降低5.4～23.7％，纯度排序为C55,H55,H50>H60>C50>C60。
40℃加速条件下放置2周，枸橼酸-枸橼酸钠缓冲体系SEC主峰含量降低0.1～2.5％，聚体含量增加0.1～0.4％，片段含量增加0～2.1％。但是所有组氨酸-组氨酸盐酸盐缓冲体系均发生明显降解反应，因此枸橼酸-枸橼酸钠缓 冲体系优于组氨酸-组氨酸盐酸体系，最佳pH为5.5。
表5.h1G4抗体缓冲体系筛选研究加速稳定性试验(40℃)数据汇总表
Figure PCTCN2021070248-appb-000007
注：N/A表示不适用。
实施例4.辅料筛选研究
本轮研究共6个备选处方(以下分别称为F1～F6)，目标蛋白h1G4抗体浓度为10.0mg/mL。缓冲体系均为20mmol/L pH 5.5的枸橼酸-枸橼酸钠溶液。除了处方F3不含氯化钠以外，其余处方中均含有0.3％氯化钠、0.02％聚山梨酯80，其中处方F1、F2和F4中甘露醇的含量分别为1％、3％和5％，单因素考察甘露醇含量对蛋白稳定性影响。对比处方F2(0.3％NaCl)及F3(0％NaCl)，考察氯化钠是否有助于增加蛋白的稳定性。F2、F5和F6处方中分别含有等渗的3％甘露醇、3％蔗糖以及1％甘氨酸，筛选有助于h1G4抗体制剂长期稳定性的最优辅料体系。h1G4抗体制剂辅料筛选研究备选处方信息见表6。
表6.h1G4抗体制剂辅料筛选研究备选处方组成
Figure PCTCN2021070248-appb-000008
取适量原液超滤换液，加辅料，调节浓度制备成如表6所示的处方，生物安全柜内采用0.22μm一次性无菌过滤器过滤样品，无菌分装至2mL西林瓶中，加13mm胶塞，轧13mm铝塑组合盖。将分装后样品放置于40℃及2～8℃的恒温药用保存箱内正置8周。于第0、1、2、4、6、8周分别取样检测。
h1G4抗体制剂辅料筛选研究单因素试验设计6个处方，根据40℃加速稳定性试验结果以及辅料优劣排序(见表7)，不含氯化钠的处方(F3)以及含有5％甘露醇的处方(F4)稳定性略差，含有1％甘露醇(F1)、3％甘露醇(F2)的处方与含有3％蔗糖的处方(F5)无显著性差异。
表7.h1G4抗体制剂辅料筛选研究高温加速试验(40℃)数据汇总表
Figure PCTCN2021070248-appb-000009
注：N/A表示不适用。*表示F4处方在40±2℃放置2周后乳光情况略重于其余处方。
实施例5.稳定剂的筛选
本轮研究通过单因素试验筛选制剂处方，共设计9个备选处方(以下分别称为F7～F15)，目标药物h1G4抗体浓度为10.0mg/mL。其中处方F7～F14的缓冲体系均为20mmol/L pH 5.5枸橼酸-枸橼酸钠，F15为20mmol/L枸橼酸钠-磷酸氢二钠，制剂辅料筛选研究备选处方信息见表8。
表8.h1G4抗体制剂辅料筛选研究备选处方组成
Figure PCTCN2021070248-appb-000010
取适量原液超滤换液，加辅料，调节浓度制备成表8处方，生物安全柜内采用0.22μm一次性无菌过滤器过滤样品，无菌分装至2mL西林瓶中，加13mm胶塞，轧13mm铝塑组合盖。将分装后样品放置于恒温摇床内40℃，200rpm振摇4周。于第0、1、2、4周分别取样，检测分析(见表9)。
表9.h1G4抗体制剂辅料筛选研究考察条件
Figure PCTCN2021070248-appb-000011
研究结果
0周时所有处方外观均为无色澄清液体，无明显可见异物，浓度在9～11mg/mL范围内，渗透压为185～540mOsmol/kg，浊度(A350)为0.006～0.009。 SEC主峰含量为98.0～98.2％，聚体含量为1.7～1.9％。CEX主峰含量为35.4～39.1％，酸性峰含量为31.6～33.8％，碱性峰含量为27.5～30.8％(见表10)。基础理化检测结果显示0周时，各处方质量良好，无明显差异。
经过最长4周的40℃加速处理，可以看出枸橼酸钠-磷酸氢二钠缓冲体系不适于h1G4抗体的稳定保存。
表10.h1G4抗体制剂辅料筛选研究振摇稳定性试验(40℃，200rpm)数据汇总表
Figure PCTCN2021070248-appb-000012
Figure PCTCN2021070248-appb-000013
注：N/A表示不适用。
结果表明，本发明的h1G4制剂成品在40℃条件下保存最长4周的情况下，外观、可见异物、蛋白质含量、SEC、CEX、pH、渗透压均未见明显变化趋势，均符合本品现行质量标准草案要求，表明所述制剂具有较好的稳定性。
应理解，在阅读了本发明的上述内容之后，本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落于本申请所附权利要求书所限定的范围。

Claims (10)

  1. 一种抗PD-1单克隆抗体的药物制剂，其特征在于，所述药物制剂包括：抗PD-1单克隆抗体、枸橼酸-枸橼酸钠缓冲液、蛋白保护剂、表面活性剂；
    所述枸橼酸-枸橼酸钠缓冲液含量为10-30mM；
    所述蛋白保护剂含量为质量/体积百分比0.5-5％，选自蔗糖、甘氨酸和甘露醇中的一种或多种，其中质量/体积比为g/L；
    所述表面活性剂为聚山梨酯80，含量为体积比0.01％-0.05％；
    所述抗PD-1单克隆抗体具有SEQ ID NO.7所示的重链可变区和SEQ ID NO.8所示的轻链可变区。
  2. 如权利要求1所述的药物制剂，其特征在于，所述药物制剂还含有等渗调节剂氯化钠，含量为质量/体积百分比0.1-0.5％，其中质量/体积比为g/L。
  3. 如权利要求1或2所述的药物制剂，其特征在于，所述药物制剂的pH值为5.0-6.5。
  4. 如权利要求1～3任一项所述的药物制剂，其特征在于，所述蛋白保护剂为甘露醇或蔗糖，其含量为质量/体积百分比1-5％，其中质量/体积比为g/L。
  5. 如权利要求1～4任一项所述的药物制剂，其特征在于，所述抗PD-1单克隆抗体的蛋白浓度为5-50mg/ml。
  6. 如权利要求1～5任一项所述的药物制剂，其特征在于，所述抗PD-1单克隆抗体是h1G4抗体，包含SEQ ID NO.10所示的重链和SEQ ID NO.9所示的轻链。
  7. 如权利要求1～6任一项所述的药物制剂，其特征在于，所述药物制剂含有：抗PD-1单克隆抗体10mg/ml，20mmol/L枸橼酸-枸橼酸钠缓冲液，质量/体积百分比为3％的甘露醇，体积百分比为0.02％的聚山梨酯80，质量/体积百分比为0.3％的氯化钠，pH 5.5，其中质量/体积比为g/L。
  8. 如权利要求1～6任一项所述的药物制剂，其特征在于，所述药物制剂含有：抗PD-1单克隆抗体10mg/ml，20mmol/L枸橼酸-枸橼酸钠缓冲液，质量/体积百分比为1-3％的蔗糖，体积百分比为0.02％的聚山梨酯80，质量/体积百分比为0.3％的氯化钠，pH 5.5，其中质量/体积比为g/L。
  9. 如权利要求1～6任一项所述的药物制剂，其特征在于，所述药物制剂含有：抗PD-1单克隆抗体10mg/ml，20mmol/L枸橼酸-枸橼酸钠缓冲液，质量/体积百分比为1％的甘氨酸，体积百分比为0.02％的聚山梨酯80，质量/体积百分比为0.3％的氯化钠，pH 5.5，其中质量/体积比为g/L。
  10. 权利要求1～9任一项所述的药物制剂在制备注射用液体制剂或冻干制剂中的应用。
PCT/CN2021/070248 2020-01-06 2021-01-05 一种细胞程序性死亡受体1抗体制剂及其用途 WO2021139642A1 (zh)

Priority Applications (8)

Application Number Priority Date Filing Date Title
JP2022541225A JP2023510229A (ja) 2020-01-06 2021-01-05 プログラム細胞死受容体１抗体製剤およびその使用
AU2021205538A AU2021205538A1 (en) 2020-01-06 2021-01-05 Programmed cell death receptor 1 antibody formulation and use thereof
KR1020227026697A KR20220124208A (ko) 2020-01-06 2021-01-05 프로그램된 세포 사멸 수용체1 항체 제제 및 그 용도
EP21738991.5A EP4088740A4 (en) 2020-01-06 2021-01-05 FORMULATION OF PROGRAMMED CELL DEATH RECEPTOR 1 ANTIBODY AND USE THEREOF
CA3162976A CA3162976A1 (en) 2020-01-06 2021-01-05 Programmed cell death receptor 1 antibody formulation and use thereof
US17/758,399 US20230048612A1 (en) 2020-01-06 2021-01-05 Programmed cell death receptor 1 antibody formulation and use thereof
BR112022012768A BR112022012768A2 (pt) 2020-01-06 2021-01-05 Formulação de anticorpo de receptor 1 de morte celular programada e uso da mesma
ZA2022/07051A ZA202207051B (en) 2020-01-06 2022-06-24 Programmed cell death receptor 1 antibody formulation and use thereof

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN202010008058.6 2020-01-06
CN202010008058.6A CN110787292B (zh) 2020-01-06 2020-01-06 一种细胞程序性死亡受体1抗体制剂及其用途

Publications (1)

Publication Number Publication Date
WO2021139642A1 true WO2021139642A1 (zh) 2021-07-15

Family

ID=69448561

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2021/070248 WO2021139642A1 (zh) 2020-01-06 2021-01-05 一种细胞程序性死亡受体1抗体制剂及其用途

Country Status (10)

Country Link
US (1) US20230048612A1 (zh)
EP (1) EP4088740A4 (zh)
JP (1) JP2023510229A (zh)
KR (1) KR20220124208A (zh)
CN (1) CN110787292B (zh)
AU (1) AU2021205538A1 (zh)
BR (1) BR112022012768A2 (zh)
CA (1) CA3162976A1 (zh)
WO (1) WO2021139642A1 (zh)
ZA (1) ZA202207051B (zh)

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110787292B (zh) * 2020-01-06 2020-04-24 上海复宏汉霖生物技术股份有限公司 一种细胞程序性死亡受体1抗体制剂及其用途
CN113797330A (zh) * 2020-06-11 2021-12-17 三生国健药业（上海）股份有限公司 一种抗pd-1单克隆抗体液体制剂
MX2022015733A (es) * 2020-06-19 2023-01-18 Sinocelltech Ltd Formulacion estable para el anticuerpo monoclonal anti-pd-1 recombinante.
MX2023001160A (es) * 2020-07-31 2023-02-22 Jiangsu Hengrui Pharmaceuticals Co Ltd Composicion farmaceutica del anticuerpo anti-pd-1 y uso del mismo.

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018052818A1 (en) 2016-09-16 2018-03-22 Henlix, Inc. Anti-pd-1 antibodies
CN109966487A (zh) * 2017-12-28 2019-07-05 上海复宏汉霖生物制药有限公司 一种包含抗pd-l1单克隆抗体的药物配制剂
CN110787292A (zh) * 2020-01-06 2020-02-14 上海复宏汉霖生物技术股份有限公司 一种细胞程序性死亡受体1抗体制剂及其用途

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106390115A (zh) * 2015-07-29 2017-02-15 上海君实生物医药科技股份有限公司 一种人源化单克隆抗体的稳定制剂
WO2018027524A1 (en) * 2016-08-09 2018-02-15 Innovent Biologics (Suzhou) Co., Ltd. Pd-1 antibody formulation
CN110354073B (zh) * 2018-04-09 2021-10-01 鲁南制药集团股份有限公司 一种免疫抑制剂单克隆抗体的液体制剂

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018052818A1 (en) 2016-09-16 2018-03-22 Henlix, Inc. Anti-pd-1 antibodies
CN109923126A (zh) * 2016-09-16 2019-06-21 上海复宏汉霖生物技术股份有限公司 抗-pd-1抗体
CN109966487A (zh) * 2017-12-28 2019-07-05 上海复宏汉霖生物制药有限公司 一种包含抗pd-l1单克隆抗体的药物配制剂
CN110787292A (zh) * 2020-01-06 2020-02-14 上海复宏汉霖生物技术股份有限公司 一种细胞程序性死亡受体1抗体制剂及其用途

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of EP4088740A4

Also Published As

Publication number Publication date
US20230048612A1 (en) 2023-02-16
CN110787292A (zh) 2020-02-14
KR20220124208A (ko) 2022-09-13
EP4088740A1 (en) 2022-11-16
ZA202207051B (en) 2023-11-29
CN110787292B (zh) 2020-04-24
CA3162976A1 (en) 2021-07-15
JP2023510229A (ja) 2023-03-13
AU2021205538A1 (en) 2022-07-14
BR112022012768A2 (pt) 2022-09-06
EP4088740A4 (en) 2024-02-21

Similar Documents

Publication Publication Date Title
WO2021139642A1 (zh) 一种细胞程序性死亡受体1抗体制剂及其用途
CA2454587C (en) Stable lyophilized pharmaceutical formulation of igg antibodies
US20220204597A1 (en) Formulations of monoclonal antibodies
RU2548772C2 (ru) Составы антитела
US20080003220A1 (en) Buffering agents for biopharmaceutical formulations
JP6429784B2 (ja) 抗体の高濃度液体製剤の製造方法
EP3215122A1 (en) Stable protein solution formulation containing high concentration of an anti-vegf antibody
KR20100016001A (ko) 항체 제제
MXPA98000684A (en) Formulation of isotonic protocols isotonic s
US20200308297A1 (en) Clinically Proven Subcutaneous Pharmaceutical Compositions Comprising Anti-CD38 Antibodies and Their Uses
JP6407174B2 (ja) 抗体の配合物および該配合物の使用
US20230250174A1 (en) Clinically Proven Subcutaneous Pharmaceutical Compositions Comprising Anti-CD38 Antibodies and Their Uses in Combination with Lenalidomide and Dexamethasone
US20200330593A1 (en) Clinically Proven Subcutaneous Pharmaceutical Compositions Comprising Anti-CD38 Antibodies and Their Uses in Combination with Bortezomib, Mephalan and Prednisone
US20230295333A1 (en) Clinically Proven Subcutaneous Pharmaceutical Compositions Comprising Anti-CD38 Antibodies and Their Uses in Combination with Pomalidomide and Dexamethasone
US20230330224A1 (en) Clinically Proven Subcutaneous Pharmaceutical Compositions Comprising Anti-CD38 Antibodies and Their Uses in Combination with Bortezomib and Dexamethasone
CN109982685A (zh) 药物配制品及其制备方法
JP4422485B2 (ja) 免疫サイトカイン含有凍結乾燥製剤
CN111375057A (zh) 一种包含抗Her2单克隆抗体的药物配制剂
US20210188941A1 (en) Stable fusion protein formulation

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 21738991

Country of ref document: EP

Kind code of ref document: A1

ENP Entry into the national phase

Ref document number: 3162976

Country of ref document: CA

ENP Entry into the national phase

Ref document number: 2022541225

Country of ref document: JP

Kind code of ref document: A

REG Reference to national code

Ref country code: BR

Ref legal event code: B01A

Ref document number: 112022012768

Country of ref document: BR

ENP Entry into the national phase

Ref document number: 2021205538

Country of ref document: AU

Date of ref document: 20210105

Kind code of ref document: A

ENP Entry into the national phase

Ref document number: 20227026697

Country of ref document: KR

Kind code of ref document: A

NENP Non-entry into the national phase

Ref country code: DE

ENP Entry into the national phase

Ref document number: 2021738991

Country of ref document: EP

Effective date: 20220808

ENP Entry into the national phase

Ref document number: 112022012768

Country of ref document: BR

Kind code of ref document: A2

Effective date: 20220627